Project Summary The incidence of obesity has increased dramatically over the last fifty years. Over half of all Americans are over-weight and over one third are classified as clinically obese. The health ramifications of this epidemic of obesity are seen in the concomitant increase in the incidence of type 2 diabetes. An estimated 29 million Americans have diabetes with 95% of those being type 2. Metformin, the most commonly prescribed therapeutic used to treat the early stages of type 2 diabetes targets the AMP-activated protein kinase (AMPK). Therefore, defining the functions and regulation of AMPK is of great significance to human health. This proposal uses baker's yeast as its model system to study the regulation of the yeast AMPK. In both yeast and human cells, the overall structure of AMPK and the modes of regulation are highly conserved. The speed and synergy of genetic and biochemical studies in yeast make this an ideal system to dissect the regulation of AMPK and apply the new knowledge to human biology. We have found that yeast AMPK regulates glucose transport by controlling which glucose transporters are maintained on the plasma membrane. We will investigate the molecular mechanism by which yeast AMPK controls the localization, function and expression of glucose transporters. In addition we have discovered a novel mechanism by which the glucose analog, 2- deoxyglucose, inhibits glucose metabolism. Yeast and cancer cells share a metabolic strategy called aerobic glycolysis that makes them extra sensitive to 2-deoxyglucose. We will use genetic, biochemical and genomic technologies to define the mechanisms underlying the toxicity and acquired resistance to this compound. Last, we have identified Snf1 serine-214 phosphorylation as a novel mechanism for the down-regulation of Snf1 signaling. We will employ a genetic screen to identify the kinase responsible for the modification.